ABSTRACT The aim of this study was to determine the

pharmacokinetics and urinary excretion patterns of the soy

isoflavones daidzein and genistein in humans. Six healthy men
with a mean age of 37 y and a mean body mass index (in kg/m
2
)
of 24 consumed a soybean flourbased meal on two occasions
6 d apart. Blood samples and total urine were collected at
intervals for the measurement of daidzein and genistein with
HPLC. Isoflavone concentrations rose slowly and reached maxi-
mum values of 3.14 0.36 mol/L at 7.42 0.74 h for daidzein
and 4.09 0.94 mol/L at 8.42 0.69 h for genistein. Elimina-
tion half-lives were 4.7 1.1 and 5.7 1.3 h for daidzein and
genistein, respectively. The slow increase in plasma concentra-
tions is consistent with the facilitation of absorption by hydroly-
sis in the small and large intestines of the glycosidic forms of the
isoflavones present in soybean-containing foods to their corre-
sponding aglycones. The rate of urinary excretion of daidzein
was greater than that of genistein throughout the postmeal peri-
od, with mean recoveries of 62 6% and 22 4% (P < 0.001)
for daidzein and genistein, respectively. However, the ratio of the
areas under the plasma concentration versus time curves for
genistein and daidzein was equal to the ratio of the concentra-
tions of the respective isoflavones in the soy meal. It is conclud-
ed that the bioavailabilities of daidzein and genistein are similar,
not withstanding the difference in urinary excretion. Am J
Clin Nutr 1998;67:86772.
KEY WORDS Soy, isoflavones, genistein, daidzein, pharma-
cokinetics, absorption, excretion, humans, urine, bioavailability
INTRODUCTION
The role of dietary flavonoids in the prevention of several
chronic diseases is the subject of intense research interest and
the soy isoflavones have been the focus of particular attention
(13). One important component of the evaluation of the poten-
tial importance of flavonoids in human health is an understand-
ing of the absorption, distribution, metabolism, and excretion of
these substances after their ingestion. However, although some
information on the pharmacokinetics and bioavailability of some
flavonoids in humans has been reported (47), much less is
known about the isoflavones and the need for more research was
highlighted recently (8).
With regard to the isoflavones, we reported previously on the
pharmacokinetics and bioavailability of daidzein (7,4-dihy-
droxy-isoflavone) and genistein (5,7,4-trihydroxy-isoflavone)
in rats (9, 10). In these studies we showed differences between
the aglycone and conjugated forms of genistein and between the
conjugates of daidzein and genistein administered as a soy
extract. In addition, the pharmacokinetics of the synthetic
isoflavone aglycone, ipriflavone (7-isopropoxy-isoflavone),
were also reported for humans (11), but relatively little is known
about the glycosidic forms of the isoflavones found in nature.
Thus, although several studies have examined urinary excretion
of daidzein and genistein (1217), plasma concentrations were
reported only at 6.5 and 24 h after single (14, 15) or multiple
(13) soy meals. In the present study, therefore, we compared the
absorption, pharmacokinetics, and urinary excretion of daidzein
and genistein in detail over 35 h after a single soybean
flourbased meal to provide further information about the possi-
ble importance of these substances to human health.
SUBJECTS AND METHODS
Subjects
The subjects were six healthy male volunteers (five white and
one Asian) aged 2148 y (mean: 37 y) with a mean body mass
index (in kg/m
2
) of 24 (range: 1939). The protocol was
approved by the CSIRO Division of the Human Nutrition Human
Ethics Committee and was fully explained to all volunteers, who
gave their informed written consent.
Study design
The study was divided into two phases spaced 6 d apart to
provide a protocol easy to manage without the capacity to obtain
blood samples over a continuous 24-h postmeal period. Subjects
were instructed to follow a soy-free diet for 1 wk before phase 1
of the study. Phase 1 was designed to determine the time
required to reach maximum plasma isoflavone concentrations.
After an overnight fast, subjects consumed a soy meal over a
period of a few minutes at 0800. The meal consisted of debit-
tered soy flour (Lowan Whole Foods, Nhill, Victoria, Australia)
that had been heat treated by the manufacturer at 105 C with
Am J Clin Nutr 1998;67:86772. Printed in USA. 1998 American Society for Clinical Nutrition
Plasma and urinary kinetics of the isoflavones daidzein and
genistein after a single soy meal in humans
1,2
Roger A King and David B Bursill
867
1
From the CSIRO Division of Human Nutrition, and the Faculty of Med-
icine, University of Adelaide, North Terrace, Adelaide, South Australia.
2
Address reprint requests to RA King, CSIRO Division of Human Nutri-
tion, PO Box 10041, Gouger Street, Adelaide, South Australia 5000. E-mail:
roger.king@dhn.csiro.au.
Received April 25, 1997.
Accepted for publication October 30, 1997.

b
y

g
u
e
s
t

o
n

A
p
r
i
l

2
1
,

2
0
1
4
a
j
c
n
.
n
u
t
r
i
t
i
o
n
.
o
r
g
D
o
w
n
l
o
a
d
e
d

f
r
o
m

moist heat during processing. The flour was suspended in
300400 mL cow milk to provide 0.84 g flour/kg body wt, to
which was added powdered chocolate flavoring to taste. This
provided 2.7 mol daidzein and 3.6 mol genistein/kg body wt.
Meals or snacks, excluding soy products, were permitted
throughout the 8-h postmeal period, but generally only liquids
were consumed until 5 h after the meal.
Ten-milliliter blood samples were taken from a medial cubital
vein into evacuated tubes containing K
2
EDTA (final concentra-
tion: 1.8 g/L) before and 1, 2, 4, 6, and 8 h after consumption of
the soy meal. Blood samples were centrifuged immediately (15
min at 4000 g and 5 C) and duplicate 200-L aliquots were
taken into screw-capped glass vials and stored at 20 C until
assayed within 4 wk. Urine was collected into bottles for 24 h
immediately before the study to provide baseline values, and for
the intervals 01, 12, 24, 46, and 68 h after the meal. Preser-
vative (1 g ascorbic acid + 1 g sodium azide) was added to the
collection bottles (18) with the pretreatment urine sample. No
preservative was added to the urine samples from the other col-
lection periods, but in all cases the samples were stored on ice
for <1 h and duplicate 200-L aliquots for isoflavone assay were
immediately taken into sealed glass vials and stored at 20 C
until assayed within 5 wk.
Phase 2 of the study was designed to determine the elimination
half-life. The meal was consumed at 2200 and blood samples
were collected 11, 12, 14, 16, 18, and 35 h after the meal. Urine
samples were collected over the periods 011, 1112, 1214,
1416, 1618, and 1835 h after the meal. Preservative was pro-
vided in the collection bottles for the 011-h and 1835-h sam-
ples. Aliquots of plasma and urine for isoflavone assay were also
taken and stored as for phase 1.
For one volunteer, blood samples were collected 1, 2, 4, 6, 8,
10, 12, 14, 16, 20, and 24 h after the 0800 meal and 0.5, 1, 2, 4, 6,
8, 11, 12, 14, 16, 18, and 35 h after the 2200 meal to validate the
two-phase design. Samples obtained during the period 18 h after
the 0800 meal and 1135 h after the 2200 meal were taken from a
medial cubital vein by using evacuated collection tubes and
processed immediately as detailed above. Corresponding urine
samples were also dispensed as aliquots immediately as detailed
above. The remaining blood samples were collected by the volun-
teer from an indwelling catheter inserted in a cephalic vein. For
these samples 5 mL blood was removed and discarded before col-
lection of a further 10 mL, which was kept on ice. Total urine sam-
ples for the corresponding intervals were also collected and stored
on ice. Blood and urine samples were transferred to the laboratory
at 0800 the next morning for immediate processing.
Analytic methods
Genistein and -glucuronidase/sulfatase (product #G0876)
were obtained from Sigma-Aldrich, Castle Hill, Australia.
Daidzein was purchased from Research Biochemicals Interna-
tional, Natick, MA. Equol was purchased from Plantech (UK),
Reading, United Kingdom. All other reagents were analytic
grade or better. The concentrations of genistein, daidzein, and
equol in HPLC standard solutions were determined by using the
extinction coefficients as described previously (9):
262
37 260,

254
27 542, and
284
4100, respectively.
The free (aglycone) and total (aglycone + glycoside)
isoflavone contents of the soy flour were measured by using the
method of Pettersson and Kiessling (19) with minor modification
as described previously (9). The total genistein content was 4.44
mol/g and the daidzein content was 3.27 mol/g, of which <
3% were in the respective free forms.
Isoflavone conjugates in plasma and urine were hydrolyzed,
extracted, and assayed by using HPLC with electrochemical
detection as described previously (9), with the following minor
modifications. Three extractions with 0.5 mL diethyl ether were
used to maximize recovery. Glass vials (Chromacol Ltd, Herts-
fordshire, United Kingdom) were used instead of plastic tubes
for incubations and ether extractions to prevent extraction from
plastic tubes of a substance that we showed would otherwise
interfere with HPLC measurement of daidzein. HPLC conditions
and equipment were as described previously (9), except that the
mobile phase consisted of 50:50:1 methanol:0.1 mol ammonium
acetate/L, pH 4.6:25 mmol EDTA/L (by vol). Typical HPLC pro-
files of plasma and urine samples collected before and after the
consumption of the soy meal are shown in Figure 1.
Recoveries for daidzein, genistein, and equol, as determined
from quadruplicate estimates from plasma and urine samples
containing known amounts, ranged from 72% to 78% with SEs
<5%. All results were corrected for the individual recovery rates.
Statistics
The significance of differences was determined by using Stu-
dents t test (INSTAT; GraphPad Software, San Diego). A P
value <0.05 was considered significant.
RESULTS
Plasma concentrations of daidzein and genistein from the sin-
gle subject in whom concentrations were continuously moni-
tored for 24 h after a soy meal at 0800 and at 2200 6 d later are
shown in Figure 2. There was a close similarity between the pro-
868 KING AND BURSILL
FIGURE 1. Typical HPLC profiles of premeal (A, C) and postmeal
(B, D) samples of urine (A, B) and plasma (C, D). d, daidzein; g, genis-
tein. Electrode response refers to the electrical response of the HPLC
electrochemical detector.

b
y

g
u
e
s
t

o
n

A
p
r
i
l

2
1
,

2
0
1
4
a
j
c
n
.
n
u
t
r
i
t
i
o
n
.
o
r
g
D
o
w
n
l
o
a
d
e
d

f
r
o
m

files obtained from the two study phases, confirming the validity
of the two-phase experimental design. Note that plasma
isoflavones were detectable as early as 0.5 h after the 2200 meal.
The validity of the two-phase design is also supported by the
data shown in Figure 3, which shows the mean plasma
isoflavone profile of the six volunteers. In particular, there was
no evidence of any discontinuity in the profile between 8 and 11
h. The mean ratio of the areas under the respective curves for
genistein and daidzein obtained by using the trapezoidal method
(20) was 1.36 0.23, which was the same as the molar ratio of
1.36 for the contents of the isoflavones in the soy flour meal.
Log transformation of the plasma isoflavone concentrations
during the period 1135 h after the meal (study phase 2) sug-
gested a single elimination rate (Figure 4). Mean correlation
coefficients for daidzein and genistein were both 0.99 (range:
0.971.00). The derived elimination half-lives (t
1/2
), together
with the mean maximum concentrations (C
max
) and the mean of
the corresponding times to reach maximum concentrations (t
max
),
are shown in Table 1. Although both t
max
and t
1/2
tended to be
higher for genistein than for daidzein, these were not signifi-
cantly different (Students unpaired t test). The higher mean C
max
value for genistein is consistent with the higher concentration of
genistein in the soy meal.
The rates of urinary excretion of daidzein, genistein, and
equol during the intervals 01, 12, 24, 46, and 68 h (phase
1), and 1112, 1214, 1416, 1618, and 1835 h (phase 2) after
the soy meal are shown in Figure 5. The mean excretion rates for
daidzein and genistein increased progressively reaching a peak at
612 h after the meal. The rate of excretion of daidzein was
greater than that for genistein during all postmeal time intervals.
Equol excretion was negligible until after the 68-h collection
period and then increased steeply. A total urine sample was also
collected during the interval 011 h after the meal for the phase
2 study, which enabled the calculation of total excretion over the
35-h period after consumption of a soy meal. The mean recover-
ies of daidzein and genistein were 62 6% and 22 4% of the
dose, respectively, which were significantly different (P < 0.001,
Students unpaired t test).
DISCUSSION
In the present study we examined in detail the concurrent pat-
terns of plasma isoflavone concentrations and urinary isoflavone
excretion after consumption of a single soy meal. In agreement
with previous studies (1216), peak rates of urinary excretion
occurred between 6 and 12 h after the meal, with more than one-
half of total excretion occurring during the first 12 h. The sub-
stantially higher urinary recovery of daidzein compared with
genistein, and the large interindividual variations in the fractions
of ingested isoflavones excreted in urine, have been uniform
observations in other studies (1217). The interindividual differ-
ences may reflect differences in gut microflora populations (13,
17) and it has been concluded (15) that the higher urinary excre-
tion of daidzein reflects a greater bioavailability of this
isoflavone. However, in the present study, the ratio of the areas
SOY ISOFLAVONE PHARMACOKINETICS IN HUMANS 869
FIGURE 2. Plasma concentrations of daidzein (A) and genistein (B)
in a subject after consumption of single soy meals at 0800 (closed sym-
bols) and 2200 (open symbols). The two meals were separated by a
washout period of 6 d during which the subject was asked not to con-
sume soybean-containing foods.
FIGURE 3. Plasma daidzein () and genistein concentrations () of
six subjects after consumption of a soy meal that provided 2.8 mol
daidzein and 3.6 mol genistein/kg body wt. Note that the data for the
time period 08 h were obtained after consumption of the meal at 0800
whereas the data for the time period 1135 h were obtained after con-
sumption of a meal at 2200 after a 6-d washout period during which sub-
jects were asked not to consume soy-containing foods. The dashed line
indicates the separation of phases 1 and 2 of the study. x

SEM.

b
y

g
u
e
s
t

o
n

A
p
r
i
l

2
1
,

2
0
1
4
a
j
c
n
.
n
u
t
r
i
t
i
o
n
.
o
r
g
D
o
w
n
l
o
a
d
e
d

f
r
o
m

under the plasma isoflavone concentration versus time curves was
the same as the ratio of the concentrations of the two isoflavones
in the soy flour meal. In agreement with this, similar plasma
daidzein and genistein concentrations were shown previously to
be accompanied by large differences in urinary recovery, but
plasma concentrations were measured only at 6.5 and 24 h after
the meal (1315) and so areas under the curves could not be cal-
culated. Similarly, consumption of isolated soy protein twice
daily for 14 d was also shown to result in a close correspondence
between the ratio of daidzein to genistein in the isolate and in
plasma obtained at 6.5 h and 7 and 14 d after the meal (21).
Unlike some other flavonoids that may be methylated, there is
no evidence that once absorbed the isoflavones undergo any
metabolic transformations other than the formation of glu-
curonide and sulfate conjugates. Thus, it is difficult not to con-
clude that the bioavailabilities of the two isoflavones were simi-
lar as was also concluded by Coward et al (21) in the study
referred to above.
The higher urinary excretion of daidzein compared with
genistein suggests a greater fractional excretion of the latter via
the bile (14, 15). Alternatively, the excretion of as yet unidenti-
fied metabolites of genistein in urine would also explain the pre-
sent results and cannot be ruled out. In rats there is significant
excretion of genistein (21, 22) and daidzein (23) in bile, although
the relative excretion of the two isoflavones in human bile and
urine is not known. However, it was shown that relatively small
differences in the structure of rutin derivatives had a major effect
on the partition of the flavonoids between urine and bile in rats
(24) and this may also be true for the isoflavones.
Notwithstanding the consistent observation of greater excretion
of daidzein than genistein in urine, there are large variations
between the different studies in the reported percentages of the
ingested isoflavones excreted. Thus, the mean recoveries of
daidzein and genistein in urine in the present study were 62% and
22% of the dose, respectively. These compare well with the data
of Lu et al (16), who reported 66.2% and 23.9% excretion for
daidzein and genistein, respectively, but are somewhat higher than
other values reported (1215), which range from 16% (13) to 49%
(14) for daidzein and from 10% (13, 15) to 16% (14) for genistein.
There are several possible explanations for this variation. First, the
nature of the soy food may inuence bioavailability through dif-
ferences in the nature of the isoavone conjugates (25, 26).
Although we did not determine the conjugation pattern in our soy
our, it is likely that it differed from those of the soy milks used
in other studies (1214, 16) because soy milk has a greater pro-
portion of simple glucosides than do less processed foods (25, 26).
These differences in conjugation pattern may inuence the ease of
hydrolysis of the glycosidic bond or bacterial degradation (27, 28)
and, hence, bioavailability. Second, in our study, care was taken to
ensure that urine samples that were obtained over a period of >2
h were collected in the presence of preservative (18), or, for sam-
ples obtained over shorter collection periods, immediately placed
on ice and rapidly subsampled and frozen. Details regarding pre-
cautions taken before freezing of samples in other studies (1216)
were not always provided; however, none reported using preserv-
ative and it is possible that variations may reect differences in
degradation during preliminary storage. The reason for this large
variation between laboratories and within laboratories is important
and warrants further study.
Log transformation of the plasma concentrations of the
isoavones over time yielded single slopes, with elimination half-
lives for daidzein and genistein of 4.7 and 5.7 h, respectively.
These values agree with the values of 2.94.4 h for daidzein and
3.86.7 h for genistein obtained indirectly by using mathematical
derivations from urinary isoavone excretion rates (12, 16). They
are also similar to those obtained for the synthetic isoavone ipri-
avone (11), but shorter than that of daidzein, one of the metabo-
lites of ipriavone, reported by the same group (11). It is possible
that the greater value for the half-life of daidzein obtained by Saito
(11) may reect generation of daidzein from ipriavone. Interest-
ingly, values for the elimination half-life of genistein similar to
those obtained in the present study were reported in rodents (9, 29,
30). However, until information is obtained regarding the extent of
biliary excretion of isoavones in humans, the signicance of this
similarity is not clear.
It has been generally accepted that dietary flavonoid glyco-
sides must be hydrolyzed to the corresponding less polar agly-
870 KING AND BURSILL
FIGURE 4. Mean plasma daidzein () and genistein () concentra-
tions (plotted on a log scale) during the period 1135 h after consump-
tion of a single soy meal that provided 2.8 mol daidzein and 3.6 mol
genistein/kg body wt. x

SEM; n = 6.
TABLE 1
Pharmacokinetic parameters of isoavones after a single soy meal
1
Isoavone t
max
C
max
t
1/2
h mol/L h
Daidzein 7.42 0.74 3.14 0.36 4.71 1.06
Genistein 8.00 0.68 4.09 0.94 5.74 1.27
1
x

SEM; n = 6. Values for t

max
, C
max
, and t
1/2
were obtained after con-
sumption of a soy meal that provided 2.8 mol daidzein and 3.6 mol gen-
estein/kg body wt. Values for t
1/2
were obtained after log transformation of
plasma concentration values during the period 1135 h after the meal.
None of the values were signicantly different (Students unpaired t test).

b
y

g
u
e
s
t

o
n

A
p
r
i
l

2
1
,

2
0
1
4
a
j
c
n
.
n
u
t
r
i
t
i
o
n
.
o
r
g
D
o
w
n
l
o
a
d
e
d

f
r
o
m

cones before significant gastrointestinal absorption can occur (7,
31, 32). However, recent evidence suggests that some flavonoids
may also be transported in their glycosidic forms (33, 34),
although the isoflavones have not been studied specifically. Sev-
eral aspects of the present study are relevant to this question. The
times required to reach peak plasma concentrations for genistein
and daidzein were 8.4 and 7.4 h, respectively, with plasma con-
centrations remaining above 50% of their maximum values for
1216 h after a single soy flour meal. The times to reach maxi-
mum plasma concentrations are much longer than the value of
1.3 h for the synthetic isoflavone aglycone, ipriflavone (11). This
suggests the need for hydrolytic cleavage of the glycosidic bond
of isoflavone glycosides of soy before significant absorption can
occur. In the present study isoflavones were detectable in plasma
30 min after the meal, and concentrations at 1 h were 40% of
their peak values. Based on the value of 3% of total isoflavones
present as aglycones in our soy meal, 6 mol daidzein was
ingested as the aglycone. The urinary excretion rate for daidzein
during the first hour showed that 2.5 mol was excreted during
this period. With use of our value of 62% of ingested daidzein
excreted in urine, the daidzein absorbed during the first hour
could be accounted for by the 3% aglycone present in the soy
meal. Similar arguments apply to genistein. For a liquid meal,
50% gastric emptying occurs within 40 min of consumption with
a median small intestinal transit time (pylorus to cecum) of 23
h (35). Thus, the rapid rise in plasma concentrations seen during
the first hour may have been due to absorption of aglycone pre-
sent in the soy meal, with the later absorption occurring after
hydrolysis of glycoside conjugates by microorganisms present in
the small and large intestines (36). Similar conclusions were
drawn in our studies in rats (9). Consistent with this conclusion
is the appearance of significant concentrations of equol in urine
only at times later than 68 h after the meal, probably reflecting
bacterial degradation of the isoflavones after entry of the meal
into the large intestine.
As discussed earlier, care should be taken in generalizing the
present results. Both bioavailability and the time course of
hydrolysis of conjugates, as well as their transport across the gut
wall and appearance in blood, may depend on the nature of the
soy food involved. It will therefore be important in future com-
parative studies to determine the influence of conjugation pat-
terns on isoflavone pharmacokinetics and bioavailability.
We thank Cherie Keatch for expert technical assistance.
REFERENCES
1. Messina MJ, Persky V, Setchell KDR, Barnes S. Soy intake and can-
cer risk: a review of the in vitro and in vivo data. Nutr Cancer
1994;21:11331.
2. Knight DC, Eden JA. A review of the clinical effects of phytoestro-
gens. Obstet Gynecol 1996;87:897904.
3. Hertog MGL, Feskens EJ, Hollman PCH, Katan MB, Kromhout D.
Dietary antioxidant flavonoids and risk of coronary heart disease:
the Zutphen elderly study. Lancet 1993;342:100711.
4. Ferry DR, Smith A, Malkhandi J, et al. Phase 1 clinical trial of the
flavonoid quercetinpharmacokinetics and evidence for in vivo
tyrosine kinase inhibition. Clin Cancer Res 1996;2:65968.
5. Gugler R, Leschik M, Dengler HJ. Disposition of quercetin in man
after single oral and intravenous doses. Eur J Clin Pharmacol
1975;9:22934.
6. Hollman PCH, Van der Gaag M, Mengelers MJB, Van Trijp JMP, de
Vries JH, Katan MB. Absorption and disposition kinetics of the
dietary antioxidant quercetin in man. Free Radic Biol Med
1996;21:7037.
7. Cova D, De Angelis L, Giavarini F, Palladini G, Perego R. Pharma-
cokinetics and metabolism of oral diosmin in healthy volunteers. Int
J Clin Pharmacol Ther Toxicol 1992;30:2933.
8. Hollman PCH. Bioavailability of flavonoids. Eur J Clin Nutr
1997;51(suppl 1):S669.
9. King RA, Broadbent JL, Head RJ. Absorption and excretion of the
soy isoflavone genistein in rats. J Nutr 1996;126:17682.
10. King RA. Daidzein conjugates are more bioavailable than genistein
conjugates in rats. Am J Clin Nutr (in press).
11. Saito AM. Pharmacokinetic study of ipriflavone (TC-80) by oral
administration in healthy male volunteers. Jpn Pharm Ther J
1985;13:722333.
12. Lu LW, Grady JJ, Marshall MV, Ramanujam VMS, Anderson KE.
Altered time course of urinary daidzein and genistein excretion dur-
ing chronic soya diet in healthy male subjects. Nutr Cancer
1995;24:31123.
13. Xu X, Harris KS, Wang H, Murphy PA, Hendrich S. Bioavailability
of soybean isoflavones depends upon gut microflora in women. J
Nutr 1995;125:230715.
14. Tew B-Y, Xu X, Wang H-J, Murphy PA, Hendrich S. A diet high in
wheat fiber decreases the biovailability of soybean isoflavones in a
single meal fed to women. J Nutr 1996;126:8717.
15. Xu X, Wang H-J, Murphy PA, Cook L, Hendrich S. Daidzein is a
more bioavailable soymilk isoflavone than is genistein in adult
women. J Nutr 1994;124:82532.
SOY ISOFLAVONE PHARMACOKINETICS IN HUMANS 871
FIGURE 5. Urinary excretion of daidzein (), genistein (), and
equol ( ) before and at intervals up to 35 h after consumption of a sin-
gle soy meal that provided 2.8 mol daidzein and 3.6 mol genistein/kg
body wt. Note that the data for the time period 08 h were obtained after
consumption of the meal at 0800 whereas the data for the time period
1135 h were obtained after consumption of a meal at 2200 after a 6-d
washout period during which subjects were asked not to consume soy-
containing foods. x

SEM; n = 6.

b
y

g
u
e
s
t

o
n

A
p
r
i
l

2
1
,

2
0
1
4
a
j
c
n
.
n
u
t
r
i
t
i
o
n
.
o
r
g
D
o
w
n
l
o
a
d
e
d

f
r
o
m

16. Lu LJW, Lin SN, Grady JJ, Nagamani M, Anderson KE. Altered
kinetics and extent of urinary daidzein and genistein excretion in
women during chronic soya exposure. Nutr Cancer
1996;26:289302.
17. Kelly GE, Nelson C, Waring MA, Joannou GE, Reeder AY. Metabo-
lites of dietary (soya) isoflavones in human urine. Clin Chim Acta
1993;223:922.
18. Adlercreutz H, Vanderwildt J, Kinzel J, et al. Lignan and
isoflavonoid conjugates in human urine. J Steroid Biochem Mol
Biol 1995;52:97103.
19. Pettersson H, Kiessling KH. Liquid chromatographic determination
of the plant estrogens coumestrol and isoflavones in animal feed. J
Assoc Off Anal Chem 1984;67:5036.
20. Chow S, Liu J. Design and analysis of bioavailability and bioequiv-
alence studies. In: Owen DB, Schucany WR, Cornell RG, Kennedy
WJ, Kshirsagar AM, Schilling EG, eds. Statistics: textbooks and
monographs. Vol 133. New York: Marcel Dekker Inc, 1992:122.
21. Coward L, Kirk M, Albin N, Barnes S. Analysis of plasma
isoflavones by reversed-phase HPLC-multiple reaction ion monitor-
ing-mass spectrometry. Clin Chim Acta 1996;247:12142.
22. Yasuda T, Mizunuma S, Kano Y, Saito K, Ohsawa K. Urinary and
biliary metabolites of genistein in rats. Biol Pharm Bull
1996;19:4137.
23. Yasuda T, Kano Y, Saito K, Ohsawa K. Urinary and biliary metabo-
lites of daidzin and daidzein in rats. Biol Pharm Bull
1994;17:136974.
24. Hackett AM, Griffiths LA. Enterohepatic cycling of O-(-hydrox-
yethyl) rutosides and their biliary metabolites in the rat. Experientia
1977;33:1613.
25. Wang H-J, Murphy PA. Isoflavone content in commercial soybean
foods. J Agric Food Chem 1994;42:166673.
26. Barnes S, Kirk M, Coward L. Isoflavones and their conjugates in
soy foodsextraction conditions and analysis by HPLC mass spec-
trometry. J Agric Food Chem 1994;42:246674.
27. Barnes S, Sfakianos J, Coward L, Kirk M. Soy isoflavones and can-
cer prevention. In: Back N, Cohen IR, Kritchevsky D, Lajtha A,
Paoletti R, eds. Advances in experimental medicine and biology. Vol
401. New York: Plenum Press, 1996:87100.
28. Farmakalidis E, Murphy PA. Isolation of 6-O-acetylgenistin and
6-O-acetyldaidzin from toasted defatted soyflakes. J Agric Food
Chem 1985;33:3859.
29. Supko JG, Malspeis L. Plasma pharmacokinetics of genistein in
mice. Int J Oncol 1995;7:84754.
30. Uckun FM, Evans WE, Forsyth CJ, et al. Biotherapy of B-cell pre-
cursor leukemia by targeting genistein to CD19-associated tyrosine
kinase. Science 1995;267:88691.
31. Setchell KDR. Non-steroidal estrogens of dietary origin: possible
roles in health and disease, metabolism and physiological effects.
Proc Nutr Soc N Z 1995;20:121.
32. Kuhnau J. The flavonoids. A class of semi-essential food compo-
nents: their role in human nutrition. In: Bigwood EJ, Cuthbertson
DP, Ganguly J, et al, eds. World review of nutrition and dietetics.
Vol 24. Basel, Switzerland: S Karger AG, 1976:11791.
33. Hollman PCH, De Vries JHM, van Leeuwen SD, Mengelers MJB,
Katan MB. Absorption of dietary quercetin glycosides and quercetin
in healthy ileostomy volunteers. Am J Clin Nutr 1995;62:127682.
34. Paganga G, Rice-Evans CA. The identification of flavonoids as gly-
cosides in human plasma. FEBS Lett 1997;401:7882.
35. Malagelada J, Robertson JS, Brown ML, et al. Intestinal transit of
solid and liquid components of a meal in health. Gastroenterology
1984;87:125563.
36. Hawkesworth G, Drasar BS, Hill MJ. Intestinal bacteria and the
hydrolysis of the glycosidic bonds. J Med Microbiol 1971;4:4519.
872 KING AND BURSILL

b
y

g
u
e
s
t

o
n

A
p
r
i
l

2
1
,

2
0
1
4
a
j
c
n
.
n
u
t
r
i
t
i
o
n
.
o
r
g
D
o
w
n
l
o
a
d
e
d

f
r
o
m